The detection of zoonotic microorganisms in Rhipicephalus sanguineus (brown dog ticks) from Vietnam and the frequency of tick infestations in owned dogs.

Dog owners are greatly concerned about tick infestations in their pets. The prevalence and dispersion of ticks and their disease-causing microorganisms have been limited from the viewpoint of dog owners in Vietnam. This study investigated the presence of tick infestation and the pathogens associated with it in canines that were brought to veterinary hospitals in Vietnam. In the survey, 1,423 dogs participated from February to October 2022. Molecular and morphological methods were utilized to identify ticks and the associated pathogens. In addition,risk variables linked to tick infestation were documented and analyzed using statistical methods. The total exposure to the brown dog tick (Rhipicephalus sanguineus sensu lato) was 29.01%. Nam Dinh has the highest tick prevalence among the research areas. Tick infestation reached its highest point between June and September in the northern region of the country, with distinct seasons showing a strong correlation with tick infestation in dogs. Out of 177 tick pools examined, 146(82.49%) tested positive for at least one infection. Mycoplasma spp. (78.53%) was the most common, followed by Anaplasma spp. (37.29%), Rickettsia felis (5.08%), Babesia vogeli, and Hepatozoon canis (2.82%). In the current study, there was a statistically significant link between tick infestation and characteristics such as age, breed, body size, lifestyle, and bathing frequency. Understanding the seasonal behavior of vector ticks is crucial for identifying individuals or animals susceptible to tick-borne diseases. Studying the distribution of ticks and their ability to carry and disseminate zoonotic germs in specific places could assist veterinarians and policymakers in implementing effective strategies to manage zoonotic infections.

The Re-Emergence of Mpox: Old Illness, Modern Challenges.

The Mpox virus (MPXV) is known to cause zoonotic disease in humans. The virus belongs to the genus Orthopoxvirus, of the family Poxviridae, and was first reported in monkeys in 1959 in Denmark and in humans in 1970 in the Congo. MPXV first appeared in the U.S. in 2003, re-emerged in 2017, and spread globally within a few years. Wild African rodents are thought to be the reservoir of MPXV. The exotic trade of animals and international travel can contribute to the spread of the Mpox virus. A phylogenetic analysis of MPXV revealed two distinct clades (Central African clade and West African clade). The smallpox vaccine shows cross-protection against MPXV infections in humans. Those who have not previously been exposed to Orthopoxvirus infections are more vulnerable to MPXV infections. Clinical manifestations in humans include fever, muscle pain, headache, and vesicle formation on the skin of infected individuals. Pathognomonic lesions include ballooning degenerations with Guarnieri-like inclusions in vesicular epithelial cells. Alterations in viral genome through genetic mutations might favor the re-emergence of a version of MPXV with enhanced virulence. As of November 2023, 92,783 cases and 171 deaths have been reported in 116 countries, representing a global public health concern. Here, we provide insights on the re-emergence of MPXV in humans. This review covers the origin, emergence, re-emergence, transmission, pathology, diagnosis, control measures, and immunomodulation of the virus, as well as clinical manifestations. Concerted efforts of health professionals and scientists are needed to prevent the disease and stop its transmission in vulnerable populations.

Protective efficacy of recombinant Toxoplasma gondii dense granule protein 15 against toxoplasmosis in C57BL/6 mice.

Toxoplasma gondii is a pervasive protozoan parasite that is responsible for significant zoonoses. A wide array of vaccines using different effector molecules of T. gondii have been studied worldwide to control toxoplasmosis. None of the existing vaccines are sufficiently effective to confer protective immunity. Among the different Toxoplasma-derived effector molecules, T. gondii dense granule protein 15 from the type II strain (GRA15 (II)) was recently characterized as an immunomodulatory molecule that induced host immunity via NF-κB. Therefore, we assessed the immunostimulatory and protective efficacy of recombinant GRA15 (II) (rGRA15) against T. gondii infection in a C57BL/6 mouse model. We observed that rGRA15 treatment increased the production of IL-12p40 from mouse peritoneal macrophages in vitro. Immunization of mice with rGRA15 induced the production of anti-TgGRA15-specific IgG, IgG1 and IgG2c antibodies. The rGRA15-sensitized spleen cells from mice inoculated with the same antigen strongly promoted spleen cell proliferation and IFN-γ production. Immunization with rGRA15 significantly enhanced the survival rate of mice and dramatically decreased parasite burden in mice challenged with the Pru (type II) strain. These results suggested that rGRA15 triggered humoral and cellular immune responses to control infection. However, all of the immunized mice died when challenged with the GRA15-deficient Pru strain or the RH (type I) strain. These results suggest that GRA15 (II)-dependent immunity plays a crucial role in protection against challenge infection with the type II strain of T. gondii. This study is the first report to show GRA15 (II) as a recombinant vaccine antigen against Toxoplasma infection.

Evaluation of Immunodiagnostic Performances of Neospora caninum Peroxiredoxin 2 (NcPrx2), Microneme 4 (NcMIC4), and Surface Antigen 1 (NcSAG1) Recombinant Proteins for Bovine Neosporosis.

Bovine neosporosis is among the main causes of abortion in cattle worldwide, causing serious economic losses in the beef and dairy industries. A highly sensitive and specific diagnostic method for the assessment of the epidemiology of the disease, as well as it surveillance and management, is imperative, due to the absence of an effective treatment or vaccine against neosporosis. In the present study, the immunodiagnostic performance of Neospora caninum peroxiredoxin 2 (NcPrx2), microneme 4 (NcMIC4), and surface antigen 1 (NcSAG1) to detect IgG antibodies against N. caninum in cattle were evaluated and compared with that of the indirect fluorescent antibody test (IFAT). The results revealed that NcSAG1 had the highest sensitivity and specificity, with values of 88.4% and 80.7%, respectively, followed by NcPrx2, with a high sensitivity of 87.0% but a low specificity of 67.0%, whereas NcMIC4 showed sensitivity and specificity of 84.1% and 78.9%, respectively, when compared with IFAT. A high degree of agreement was observed for NcSAG1 (k = 0.713) recombinant protein, showing the highest diagnostic capability, followed by NcMIC4 (k = 0.64) and NcPrx2 (k = 0.558). The present study demonstrates that NcSAG1 is helpful as an antigen marker and also demonstrates the potential immunodiagnostic capabilities of NcPrx2 and NcMIC4, which could serve as alternative diagnostic markers for detecting N. caninum infection in cattle. These markers may find utility in future treatment management, surveillance, and risk assessment of neosporosis in livestock or other animal host species. Further research should be directed toward understanding the in vivo immune response differences resulting from immunization with both recombinant proteins.

Prevalence of gastrointestinal parasitic infections in wild mammals of a safari park and a zoo in Bangladesh.

In safari parks and zoos, wild animals are kept mainly for recreational purposes. Animals in these enclosures are also crucial for the education, research, and conservation aspect. To ensure better management and good health of wild animals in captivity, it is essential to monitor the occurrence of gastrointestinal (GI) parasitic (helminths and protozoa) infections. The current investigation was undertaken to investigate the prevalence of GI parasitic infections in wild mammals at Bangabandhu Sheikh Mujib (BSM) safari park and Chattogram (CTG) zoo of Bangladesh. A total of 72 individual faecal samples were collected from 25 species of wild mammals. Routine qualitative (e.g. direct smear, sedimentation, and flotation) and quantitative (e.g. McMaster technique) tests were performed to identify the eggs or oocysts of helminths and protozoa. Results demonstrated that wild mammals of both BSM safari park and CTG zoo were infected with a total of 17 genera/species of helminths and protozoa. The overall prevalence of GI parasitic infections in wild mammals of both zoological parks was 65.3% (95% confidence interval [CI]: 53.14-76.12), whereas it was 72.4% (95% CI: 52.76-87.27) in the BSM safari park and 60.5% (95% CI: 44.41-75.02) in the CTG zoo. In both zoological parks, infection with nematodes was more frequent compared to other helminth into the wild mammals. The herbivores were more infected with GI parasites than carnivores and omnivores of both BSM safari park and CTG zoo. The mean eggs/oocysts per gram of faeces was the highest in the carnivores compared to herbivores and omnivores of both enclosures. The findings of the current study demonstrated that wild mammals of both BSM safari park and CTG zoo suffered from various GI parasitic infections. Regular monitoring along with proper therapeutic measures may reduce the severe consequences of GI parasitic infections in captive wild animals.

First report of knowledge and practices towards toxoplasmosis among cat owners: A cross-sectional survey in Bangladesh.

Raising awareness about Toxoplasma gondii infection among cat owners in Bangladesh is indispensable to formulate persuasive management tactics to avoid zoonotic infections from pet cats. However, to the authors' best knowledge, no studies have been performed in Bangladesh to determine knowledge and practices of toxoplasmosis in cat owners. Therefore, the objectives of the current study were to cover this research gap. We carried out a cross-sectional study in Bangladesh from June 2020 through December 2021. A structured online questionnaire was distributed to cat owners, which were voluntarily completed by them. The questionnaire included socio-demographic data, aetiology, transmissions, clinical signs, and preventive practices towards toxoplasmosis. Overall, 1,019 cat owners participated voluntarily in the cross-sectional survey. Among them, 793 (77.82%) participants showed poor knowledge regarding toxoplasmosis. Under specific knowledge sections, 62.51% of the participants revealed incorrect knowledge that toxoplasmosis was a zoonotic disease. In the same way, (72.03-85.77) % of the cat owners were unaware that the disease could be transmitted from improperly washed vegetables, raw or undercooked meat and fish, and contaminated water and milk with cat faeces. Respondents' age, education, occupation, residence type, and marital status were significantly (p < .05) associated with their knowledge level. Besides, 94.11% of cat owners had a good practice level. They followed good practices in different issues; however, they practiced those activities without knowing their impacts on disease control. Cat owners' age, education, occupation, and residence type had a significant (p < .05) association with the practice level against toxoplasmosis. This is the first study highlighting the low level of knowledge among cat owners about toxoplasmosis in Bangladesh. These knowledge gaps could increase the risk and transmission of Toxoplasma gondii infection among them and their families. The survey recommends the arrangement of educational training and programmes to increase the awareness of toxoplasmosis among cat owners.

Prevalence of helminth and protozoan infections in pet birds of Chattogram, Bangladesh.

BACKGROUND: Parasitic diseases such as helminths and protozoa are considered one of the major impediments in the rearing of pet birds. The current study was undertaken to determine the prevalence of helminths and protozoal infections in different captive pet birds in Chattogram metropolitan area, Bangladesh. METHODS: A total of 549 pooled faecal and 311 individual blood samples were collected from different species of pet birds during June 2019 to May 2020. The faecal samples were examined following routine microscopic tests to identify the eggs and oocysts of helminths and protozoan parasites, respectively. Polymerase chain reaction (PCR) was performed to determine the haemoprotozoan parasites. RESULTS: The prevalence of helminth infestations in pet birds was 8.01% (95% confidence interval [CI]: 5.88-10.61), where infestation caused by nematodes was the highest (7.47%, CI: 5.41-10). Ascaridia and Capillaria spp. infestations were the commonest helminths recorded in different groups of pet birds. The overall prevalence of intestinal protozoal infections was 11.11% (CI: 8.61-14.04) in pet birds. The most commonly occurring protozoal infections were Eimeria spp. (7.83%, CI: 5.73-10.41) followed by Isospora spp. (2.91%, CI: 1.67-4.69). The overall prevalence of haemoprotozoan parasites was 2.25% (CI: 0.91-4.58) in different groups of pet birds. The highest prevalence was recorded in Plasmodium spp. (1.29%, CI: 0.35-3.26) followed by Leucocytozoon and Haemoproteus spp. CONCLUSIONS: The data generated in this study are the first of its type, which would be beneficial to the veterinary practice, aviculturists, pet bird owners and breeders in Bangladesh to respond appropriately for the prevention and control of the disease.

Neospora caninum surface antigen 1 is a major determinant of the pathogenesis of neosporosis in nonpregnant and pregnant mice.

Introduction: NcSAG1 is one of most widely investigated antigens of Neospora caninum in various research fields. Such studies demonstrated the proficiency of NcSAG1 in the regulatory process of parasite adhesion and invasion of host cells. Accordingly, the contribution of NcSAG1 to the pathogenesis of neosporosis can undoubtedly be extrapolated, but direct evidence is lacking. Herein, we provide the first successful attempt at the gene disruption of NcSAG1 and novel data on the invasion and virulence potentials of N. caninum in vitro and in vivo. Methods: The disruption of the NcSAG1 gene was applied using the CRISPR/Cas9 system and confirmed by PCR, western blot and indirect fluorescent antibody tests as NcSAG1 knockout parasites (NcSAG1KO). Then, we investigated the role of NcSAG1 in the growth kinetics of the parasite in vitro. Results and discussion: The deletion of the NcSAG1 gene significantly decreased the infection rate and reduced the egress rate of the parasite. An in vivo study using nonpregnant female and male BALB/c mice revealed a significantly higher survival rate and lower body weight change in the group infected with the NcSAG1KO parasite than in the parental strain (Nc-1)-infected group. Regarding the vertical transmission model of BALB/c mice, the absence of the NcSAG1 gene significantly enhanced the survival of pups and greatly lowered the parasite burden in the brains of pups. In conclusion, our study suggested NcSAG1 as a key molecule in the pathogenesis of N. caninum.

Seroprevalence of Toxoplasma gondii infection in sheep and goats from different geographical regions of the world: Systematic review and meta-analysis.

The Toxoplasma gondii is an obligate intracellular protozoan parasite which significantly impact small ruminant productivity, international animal trade and transboundary movement of animal across the globe. The seroprevalence of T. gondii infection (toxoplasmosis) in sheep and goats is widely studied in many parts of the world and there is a lack of comprehensive information on prevalence estimates considering the global and regional perspectives. The aim of the study was to use systematic review and meta-analysis methods to estimate the global and regional pooled seroprevalence of T. gondii infection in sheep and goats, as well as factors that influence prevalence estimations. Relevant articles reporting the seroprevalence of toxoplasmosis in sheep and/goats were searched in five electronic databases, including PubMed, Web of Science, Scopus, Embase and ProQuest. After the publications were checked to verify they fit the inclusion criteria, a total of 225 articles were included in the systematic review and meta-analysis, reflecting data from 70 countries/regions. The pooled prevalence was estimated using a random effect meta-analysis model. Overall, the seroprevalence of T. gondii infection was 33.86% (95% CI: 30.47-37.25%) in sheep and 31.78% (95% CI: 28.99-34.58%) in goats, with significant variation in prevalence estimates across geographical locations (p < .001). Substantial heterogeneity (I2  > 75%) was observed in most pooled seroprevalence estimates. The T. gondii infection in global sheep and goat population showed uptrend over the period. This information would be useful for epidemiologist, health authorities and farmers in order to plan future T. gondii survey and infection management strategies both locally and internationally.

Advances in vaccine development and the immune response against toxoplasmosis in sheep and goats.

Toxoplasmosis is a zoonotic, parasitic infection caused by the intracellular, apicomplexan parasite Toxoplasma gondii, which infects all homeothermic animals including humans. The parasite has a major economic impact on the livestock industry. This is especially true for small ruminants (sheep, goats) as it is one of the most likely reasons for reproductive disorders in these animals. Primary infection in sheep and goats can result in a fetus that is mummified or macerated, fetal embryonic death, abortion, stillbirth, or the postnatal death of neonates, all of which threaten sheep and goat rearing globally. Humans can also become infected by ingesting bradyzoite-containing chevon or mutton, or the contaminated milk of sheep or goats, highlighting the zoonotic significance of this parasite. This article reviews the advances in vaccine development over recent decades and our current understanding of the immune response to toxoplasmosis in small ruminants (sheep, and goats).

Molecular detection of Toxoplasma gondii in aborted fetuses of goats in Chattogram, Bangladesh.

BACKGROUND AND AIM: Toxoplasma gondii is a protozoan parasite that is responsible for the major cause of congenital diseases, abortion, and stillbirth in humans and farm animals. Primary infection in pregnant goats due to T. gondii leads to abortion and significant economic losses in the livestock industry. Moreover, very few studies have been performed for the identification of T. gondii from aborted fetuses of goats. The study was conducted for the molecular identification of Toxoplasma gondii from aborted fetuses of goats in Chattogram, Bangladesh. MATERIALS AND METHODS: Twenty aborted fetuses of goats were collected from 52 farms in the study area. A nested polymerase chain reaction (PCR) assay targeting the B1 gene was performed, and a positive sample yield of 197 bp amplified DNA products consistent with T. gondii. RESULTS: The overall prevalence of toxoplasmosis in the aborted fetus of goats was 35%. Heart muscle, liver, brain, and placenta showed positive PCR results. The risk factors related to the does age, presence of cats in farms, and aborted fetus age were found to be statistically significant (p<0.05). Our results showed that T. gondii is a major possible causal factor for abortion and reproductive failure in goats. The high prevalence of T. gondii infection in aborted fetuses of goats revealed that T. gondii could be imperative in causing reproductive failure in goats. CONCLUSION: Active or congenital toxoplasmosis was shown by the presence of T. gondii in fetal tissues, which is a matter of concern as this parasite has zoonotic significance and causes economic hazards to the livestock industry by causing various reproductive problems. Therefore, proper control measures and strategies are needed to reduce the rate of abortion in goats, ultimately saving the livestock industry.

Toxoplasma gondii GRA15 DNA Vaccine with a Liposomal Nanocarrier Composed of an SS-Cleavable and pH-Activated Lipid-like Material Induces Protective Immunity against Toxoplasmosis in Mice.

Toxoplasma gondii affects the health of humans and livestock and causes severe illness in the fetus and immunocompromised individuals. Because of the high incidence and severe consequences of T. gondii infection, a safe and suitable vaccine is needed. We found that lipid nanoparticles (LNPs) consisting of a series of functional materials prepared with vitamin E, such as SS-cleavable and pH-activated lipid-like materials (ssPalmE), were a safe and efficient way to develop next-generation DNA vaccines. In this study, we prepared ssPalmE-LNP to encapsulate pCpG-free-T. gondii dense granule protein 15 DNA (ssPalmE-LNPTgGRA15). Following a challenge infection with avirulent PLK strain of T. gondii, the mice immunized with ssPalmE-LNPTgGRA15 had a significantly higher survival rate and lower clinical scores compared with unimmunized and ssPalmE-LNPnon-coding-immunized mice. Immunization of mice with the ssPalmE-LNPTgGRA15 led to a significantly higher production of specific IgG1 and IG2c antibodies compared with unimmunized and ssPalmE-LNPnon-coding-immunized mice, while there was no statistically significant difference in the concentration of serum interferon-gamma at the acute stage of the infection. These findings indicate that ssPalmE-LNP is an effective cargo for the transportation of DNA vaccines for protozoan infections. To explore the mechanism of protective immunity induced by ssPalmE-LNPTgGRA15, further immunological study is needed in the future.